IS 12572 ( Part 3 ) : 1988 Indian Standard GUIDE FOR EVALUATION OF MEDICAL DEVICES FOR BIOLOGICAL HAZARDS PART 3 METHOD OF TESTING BY TISSUE IMPLANTATION Ymh m-w ( Reaffirmed 2006 ) UDC 615.3 : 614.8 : 616-002 : 616-069-843 DESCRIPTORS : MEDICAL DEVICES, BIQLO;;ICAL HA7ARDS TISSUE IMPLANTATION BtJREAU O F I N D I A N STAXDARDS 9 BAHADI;R StiAti ZAFAR MAKci NEW DELI11 t l(Jtr()i' Priw (;roup 2 MANAL BI IRVAN, J14(17 1989 Medical Glass Instruments and Appliances Sectional Committee, CPDC 12 FOREWORD This Indian Standard ( Part 3 ) was adopted by the Bureau of Indian Standards on 14 September 1988, after the draft finalized by the Medical Glass Instruments and Appliances Sectional Committee had been approved by the Consumer Products and Medical Instruments Division Council. This part ( Part 3 ) is one of a series of methods of test for assessing the biological hazards of medical devices that directly come in contact with body tissues or substances that are to be introduced into the body by means other than oral. Part 2 of this standard provides guidence on the selection of recommended methods of test for the assessment of biological hazards in medical devices which has been referred to in this part. This standard is proposed to be published in 12 parts. Other parts are as follows: Part I Terminology Part 2 Selection of biological methods of test Part 4 Method of test for systemic toxicity : Assessment of acute toxicity of extracts from medical devices P:irt 5 Part 6 Method of test for intracutancous reactivity of extracts from medical devices Method of test for systemic toxicity : Assessment of pyrogenicity in rabbits of extracts from medical devices Part 7 Methods of test for sensitization : Assessment of potential of medical devices to produce delayed contact dermatitis Part 8 Part 9 Method of test for skin irritation of extracts from medical devices Method of test for skin irritation of solid medical devices Part 10 Method of test for dental materials Part I1 Method of test for eye irritation Part 12 Methods of test for toxicity to ceils in culture of extracts from medical dcviccs In the preparation of this standard, assistance has been derived from BS 5736 : Part 2 : 1981 `Evaluation of medical devices for biological hazards: Part 2 Methods of testing by tissue implantation', issued by the British Standards Institution, IJK. IS 12572 ( Part 3 ) : 1988 Indian Standard GUIDEFOREVALUATIONOFMEDICAL DEVICESFORBIOLOGICALHAZARDS PART 3 METHOD OF TESTING BY TISSUE IMPLANTATION 2.6 Positive Control Specimen A piece of material which, when implanted by the procedure described in 5 produces a positive reaction (.yep 7.4). NOTE - Tin stabilized polyvinylchloride is s u i t able. I SCOPE 1.1 This standard ( Part 3 ) describes a method of test designed to provide information on the effects of direct contact of a test material with living tissue when implanted into the paravertebral muscle of the rabbit for a period of 7 days. 1.2 This method of test is recommended in the initial assessment of devices in category A and C ( .W Part 2 of this standard ) : a) Cutegor_r A - Those intended for long-term (long-term covers a period from a few months to permanent use) implantation within the body tissue, for example, artery grafts and hip prostheses; and b) Cutegory C - Those intended for shortterm (short-term covers a period from a few minutes to several weeks continuous use) use within the body or in contact with mucosai surfaces, for example, tracheal tubes, urinary catheters and intravenous cannulac. 2 DEFINITIONS For the purpose of this standard. the following definitions shall apply. 2.1 Final Product The medical devices in its ready-for-use state. 2.2 Implant The test spccimcn, positive control specimen or negative control specimen (see 2.8, 2.5 and 2.6) that has been implanted. 2.3 Implant Site The implant and 5 mm tissue surrounding it, measured from the centrc of the implant. 2.4 Medical Device Any item used in medical treatment, diagnosis or contraception, not intended to have a phsfmacological action on the body. 2.5 Negative Control Specimen A piece of m:Iterial which, when implanted by the p r o c e d u r e described in 5, PI-odwxs a n e g a t i v e reaction (.W 7.3). NOTE - Additive-l'l,ce pc>lyeth,vlenc is suitable. 2.7 Test Material The final product or sample of final product that is to be tested. 2.8 Test Specimen The piece of test material that is implanted. 3 ANIMALS AND HUSBANDRY 3.1 Not fewer than three rabbits of white strain, preferably Newzeal and type, shall be used. They shall be healthy adults whose paravertcbral muscles are sufficiently large in size to allow for implantation of test and control specimens as described in 5.2. 3.2 The annnals shall be housed individually and shall have free access lo food and water. 4 TEST AND CONTROL SPECIMENS 4.1 Number of Specimens Required The mmimum number of specimens for Implantation in each rabbit iI1 accordance with 5 shall bc: a) not fewer than two positive control s ecimens, b) not fewer than two negative control specimens, and c) not fewer than four test specimens. may b e n e c e s s a r y t o i m p l a n t more specimens than the minimum rcqtlired because 01 l o s s , I'OI c\tample, by extlusion. of implants dul-ing t h r 7 da> test pcriotl. NOTE -- It 4.2 Sterilization and Handling of Specimens 4.2.1 The specimens frc>rn prc-sterilized devices and the pre-stcrlli/ed c o n t r o l s shall hc axptically handled. I IS 12572 ( Part 3 ) : 1988 4.3 Preparation of Specimens Immediately before implanting, cut or shape the test materials and, if necessary, the controls, into specimens nominally JO x I x I mm with smooth sides to minimize mechanical trauma during implantation. 4.3.1 Place each specimen in a sterile solution containing 9 g/l sodium chloride. 5. TEST PROCEDURE 5.1 Preparation of Animals 5.1.1 Anaesthetize the rabbits by injection, for example, with 30 mg/kg pentobarbetone sodium intravenously, and record the anaesthetic and the route of administration used. 5.1.2 Clip the fur on the back of the rabbits close to the skin and swab the clipped area with nntiseptic solulion. 5.2 Implarltation of Specimens 5.2.1 Implant in one of the rabbits, the appropriate number of test and control specimens in the positions shown in Fig. 1. using the procedure described in 5.2.3 to 5.2.7. 5.2.2 Each implant shall be at least 10 mm from any other implant and the positive control specimen shall be at Jeast 30 mm to the rear of other implants. 5.2.3 Aseptically place a specimen into a sterile needle ( hypodermic type ) of cannular length I9 to 35 mm and of a bore such that the specimen is a snug but not tight fit, for example, I'8 mm bore. 5.2.4 Insert a sterile stylet behind the specimen in the needle. 5.2.5 Insert the needle vertically into the paravertebral muscle of the rabbit. 5.2.6 Hold the stylet in place whilst the needle is gently withdrawn leaving the stylet in place. 5.2.7 When the needle is free from the skin, withdraw the stylet likewise, leaving the specimen implanted within the muscle. 5.2.8 Repeat the implantations in two filrther rabbits. 5.3 Recovery of Implant Sites 5.3.1 After the implants have been in position for 7 days, kill the rabbits with an overdose of anaesthetic and place them in the prone position with the legs splayed. 5.3.2 Carefully excise the implant sites, leaving the implants in position, lEST SPECIMENS- -NEGATIVE CONTROL SPECIMENS -POSITIVE CONTROt SPECIMENS TAIL IS 12572 ( Part 3 ) : 1 9 8 8 5.4 Examination of Implant Sites 5.4.1 Microscopic Examination 5.4.1.1 Examine each excised implant site under normal vision or with the aid of a low magnification lens. Record the nature, extent and distribution of any tissue reaction observed. 5.4.1.2 If any negative or positive control specimen evokes a reaction other than that described in 7.3 and 7.4 respectively, the results of the test specimens in that rabbit shall be rejected and the test repeated in a further rabbit. 5.4.1.3 If any test specimen implant site shows a p Gtivc reaction ( see 7.4 ), at least one of these sites shall be selected for histological examination ( See 5.4.2 ) to confirm the response. 5.4.1.4 If any test specimen implant sites shows a negative reaction ( see 7.3 ), all the test specimen implant sites and the negative control implant sites shall be recovered for histological examination to confirm the response. 5.4.2 Histologicnl Examination 5.4.2,l Preserve the excised implant sites specified in 5.4.1 in formal saline. 5.4.2.2 Prepare sections transverse to the excised implant sites. O T E- I d e a l l y , the implanf s h o u l d r e m a i n i n place during preparation for sectioning to ensure correct orientation of the surrounding tissue unless adverse reaction with dehydrating or defatting solvents i< likely to occur. Hard implants may be removed bcforr cutting of sections, if cutting would othcrwixe he difiicult. 5.4.2.4 Exatnine the histological sections microscopically and record the findings. 6. PRESENTATION OF RESULTS 6.1 The results shall be submitted in a test report that includes a complete record of the procedures and anv other relevant data used in the assessment of results as described in 7. 6.2 If more than the minimum number of test specimens or control specimens are recovered ( see 4.1 ), all recovered specimens shall be considered as part of the test. 7. ASSESSMENT OF RESULTS 7.1 The assessment of results should be carried out by a toxicologist who is aware of the conditions of use for the final product and has appropriate chemical and biological data concerning it [ see 3.3 and 3.4 of IS 12572 ( Part 2 ) : 1988 1. 7.2 The overall assessment of the test results shall be carried out by taking into consideration all information in the test report. 7.3 A reaction shall be considered a negative reaction if there is no reaction or if there is a reaction that can be attributed to experimental trauma, typically asymmetrical, non-necrotic and non-inflammatory, 7.4 A reaction shall he considered a positive reaction if there is a toxic reaction, typically with necrosis and inflammation s~~mmetrically around the implant. 7.5 A test material shall have evoked a positive response if any test specimen obtained from it evokes a positive rcnction. 7.6 The results of the assessment shall be recorded in the test report. N 5.4.2.3 Stain the sections with haemotoxylin a n d eosin. 3 Standard Mark The use of the Standard Mark is governed by the provisions of the Bureau of Indian Standards Act, 1986 and the Rules and Regulations made thereunder. The Standard Mark on products covered by an Indian Standard conveys the assurance that they have been produced to comply with the requirements of that standard under a well defined system of inspection, testing and quality control which is devised and supervised by BIS and operated by the producer. Standard marked products are also continuously checked by BIS for conformity to that standard as a further safeguard. 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